%0 Journal Article %T Production of Recombinant Human Granulocyte-Colony Stimulating Factor by Pichia pastoris %J Iranian Journal of Biotechnology %I National Institute of Genetic Engineering and Biotechnology of Iran %Z 1728-3043 %A Bahrami, Ali %A Shojaosadati, Seyed Abbas %A Khalilzadeh, Rasoul %A Saeedinia, Ali Reza %A Vasheghani Farahani, Ebrahim %A Mohammadian-Mosaabadi, Jafar %D 2007 %\ 07/01/2007 %V 5 %N 3 %P 162-169 %! Production of Recombinant Human Granulocyte-Colony Stimulating Factor by Pichia pastoris %K Pichia pastoris %K rhG-CSF %K Fed-batch culture %K Methanol %R %X Human granulocyte-colony stimulating factor (hG-CSF) cDNA was expressed in the methylotrophic yeast Pichia pastoris under the control of the alcohol oxidase (AOX1) promoter. An expression vector for hG-CSF secretion was constructed using vector pPIC9. Higher levels of hG-CSF was obtained using a P. pastoris Mut+ (methanol utilization fast) phenotype. The effects of environmental factors such as temperature and pH on the P. pastoris cell growth and hG-CSF production during fermentation were investigated. Cell growth and hG-CSF production were found to be optimal at 28°C and pH 6.0. A fed-batch fermentation process was also developed to obtain high cell density and higher levels of protein expression. Using a high cell density cultivation method, cell dry weight and hG-CSF concentration reached 100 g/l and 35 mg/l, respectively. %U https://www.ijbiotech.com/article_7021_0046d351d8223494763708302c415dc7.pdf